We describe the identification of Leishmania infantum infection in Eleonora from Toledo (1522–1562), wife of Cosimo I de’ Medici and member of one of the major political Italian families during the Renaissance. The positive identifi cation of Leishmania infection was achieved in bone samples by 2 independent approaches. First, a molecular ancient DNA (aDNA) analysis identifi ed a specific 123-bp fragment of a conserved region of the minicircle molecule of the parasite´s kinetoplastid mitochondrial DNA, which on direct sequencing showed a Leishmania-specific sequence compatible with L. infantum (see online Appendix Figure, wwwnc.cdc.gov/EID/article/18/1/10-2001-FA1.htm). This PCR result was independently replicated in 2 laboratories and additionally supported by the second approach, a protein assay showing a concomitant positive reaction by detecting IgG against L. infantum by Western blot sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Direct sequencing of the Leishmania aDNA identifi ed a strain with high homology to L. infantum. Accordingly, we obtained a 98% concordance rate between our sequence and that of L. infantum (expect rate 6e-47, identity rate 113/118) (online Appendix Figure). The rates for other Leishmania species indicated that concordance for those species was less probable. For the protein assay, fractionated proteins from a lysate of late-logphase promastigotes of L. infantum ZMON-1 (World Health Organization code MHOM/TN/1980/IPT-1) were electroblotted onto nitrocellulose membrane, and antibody detection was conducted on a Bio-Rad (Hercules,CA, USA). Multiscreen apparatus (6). Antibodies against L. infantum selectively reacted in a supernatant of protein extract from Eleonora, thereby confi rming the immunologic identifi cation of the protozoal infection. The response of IgG against L. infantum whole-parasite antigens revealed specific recognition of 8 polypeptides ranging from 14–16 kD to 184 kD. This pattern of bands is consistent with a symptomatic form of VL as shown by the 14 to 16–kD bands. Our molecular and serologic identifi cation of Leishmania infection in a historically prominent person from southern Europe has major relevance. This information might be useful for monitoring the infection and its pathogen throughout history and might provide data on the host–pathogen interaction over different periods.
Visceral Leishmaniasis during Italian Renaissance, 1522-1562
GIUFFRA, VALENTINA;FORNACIARI, GINO
2012-01-01
Abstract
We describe the identification of Leishmania infantum infection in Eleonora from Toledo (1522–1562), wife of Cosimo I de’ Medici and member of one of the major political Italian families during the Renaissance. The positive identifi cation of Leishmania infection was achieved in bone samples by 2 independent approaches. First, a molecular ancient DNA (aDNA) analysis identifi ed a specific 123-bp fragment of a conserved region of the minicircle molecule of the parasite´s kinetoplastid mitochondrial DNA, which on direct sequencing showed a Leishmania-specific sequence compatible with L. infantum (see online Appendix Figure, wwwnc.cdc.gov/EID/article/18/1/10-2001-FA1.htm). This PCR result was independently replicated in 2 laboratories and additionally supported by the second approach, a protein assay showing a concomitant positive reaction by detecting IgG against L. infantum by Western blot sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Direct sequencing of the Leishmania aDNA identifi ed a strain with high homology to L. infantum. Accordingly, we obtained a 98% concordance rate between our sequence and that of L. infantum (expect rate 6e-47, identity rate 113/118) (online Appendix Figure). The rates for other Leishmania species indicated that concordance for those species was less probable. For the protein assay, fractionated proteins from a lysate of late-logphase promastigotes of L. infantum ZMON-1 (World Health Organization code MHOM/TN/1980/IPT-1) were electroblotted onto nitrocellulose membrane, and antibody detection was conducted on a Bio-Rad (Hercules,CA, USA). Multiscreen apparatus (6). Antibodies against L. infantum selectively reacted in a supernatant of protein extract from Eleonora, thereby confi rming the immunologic identifi cation of the protozoal infection. The response of IgG against L. infantum whole-parasite antigens revealed specific recognition of 8 polypeptides ranging from 14–16 kD to 184 kD. This pattern of bands is consistent with a symptomatic form of VL as shown by the 14 to 16–kD bands. Our molecular and serologic identifi cation of Leishmania infection in a historically prominent person from southern Europe has major relevance. This information might be useful for monitoring the infection and its pathogen throughout history and might provide data on the host–pathogen interaction over different periods.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
